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Promega renilla control plasmid phrl-tk
Renilla Control Plasmid Phrl Tk, supplied by Promega, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/phrl-tk+control+plasmid/pgl3+basic/pm40344170-326-27-31
Average 90 stars, based on 1 article reviews
renilla control plasmid phrl-tk - by Bioz Stars, 2026-10
90/100 stars

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Related Articles

Transfection:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Plasmid Preparation:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Control:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Expressing:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Luciferase:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Mutagenesis:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Activation Assay:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Labeling:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Activity Assay:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Immunoprecipitation:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Incubation:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Western Blot:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Translocation Assay:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Fluorescence:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).

Construct:

Article Title: WT1-mediated gene regulation in early urogenital ridge development.
Article Snippet: The Wilms tumor protein WT1 is involved in the development of several organs, including the gonads.. WT1 mutations in humans lead to syndromes associated with impaired sexual development and Wt1 knockout mice show regression of gonad anlagen.. As a transcription factor, WT1 fulfills its function by regulating a set of target genes.

Article Title: Thyroid Hormone Targets Matrix Gla Protein Gene Associated With Vascular Smooth Muscle Calcification
Article Snippet: RAOSMCs in the contractile state, grown on a 24-well plate, were transiently cotransfected with 225 ng/well of each promoter luciferase plasmid and 75 ng/well of phRL-TK control plasmid (Promega) using FuGene6 (Roche).

Article Title: Ligand induced dissociation of the AR homodimer precedes AR monomer translocation to the nucleus
Article Snippet: PC-3 cells were transfected with reporter plasmid (pGL3), phRL-TK control plasmid (Promega) and pcDNA3.1 expression plasmid for ARs or GRs with Lipofectamine 2000 and treated with ligands in the serum free media for 24 h. Cell lysate was subjected to Dual Luciferase Assay System (Promega).

Article Title: Wilms' Tumor Protein Wt1 Is an Activator of the Anti-M?llerian Hormone Receptor Gene Amhr2
Article Snippet: Cells were split at 5 × 10 4 to 8 × 10 4 TM4 cells or 2 × 10 4 to 3 × 10 4 M15 cells per well and transfected 24 h later with 750 ng expression construct, 200 ng promoter reporter construct, and 50 ng phRL-Tk control plasmid (Promega).

Article Title: Post-transcriptional downregulation of sarcolipin mRNA by triiodothyronine in the atrial myocardium.
Article Snippet: Neonatal rat atrial myocytes were incubated with serum-free medium for 24 h and then they were transiently co-transfected with 300 ng of each SLN promoter luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega), using FuGene 6 (Roche).

Article Title: A Human Phospholamban Promoter Polymorphism in Dilated Cardiomyopathy Alters Transcriptional Regulation by Glucocorticoids
Article Snippet: For promoter-reporter studies, after 24 hr incubation with serum-free medium, the myocytes were transiently cotransfected with 300 ng of each PLN luciferase test plasmid and 75 ng of phRL-TK control plasmid (Promega).



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( A ) Averaged fura-2 responses, their peak amplitude (middle graph bar) and slope (right graph bar) evoked by Ca 2+ re-addition in Tg treated (1 µM, 8 min) Jurkat cells lines generated by CRISPR with control or ORAI1-targeted guiding sequences and stably re-expressing either WT or C143A ORAI1-YFP. Data are mean ± SEM of 210 (Control), 242 (KO), 189 (WT), and 203 (C143A) cells from three independent experiments ( B ) Western blot showing the amount of biotinylated GFP immunoreactivity in the PM in Jurkat CRISPR ORAI1 cells reconstituted with WT or C143A ORAI1-YFP. Representative of 2 independent experiments. ( C ) Individual (thin lines) and averaged (thick line) fura-2 recordings of Jurkat CRISPR ORAI1 cells reconstituted with WT or C143A ORAI1-YFP, exposed to CD3/CD28-coated beads in Ca 2+ containing solution (left). Averaged peak and integrated responses evoked by CD3/CD28 beads in individual cells during the recording period (right). Data are from 52 cells (WT) and 72 cells (C143A) from three independent experiments. ( D ) Relative changes <t>in</t> <t>NFATC-Luciferase</t> vs. housekeeping- <t>Renilla</t> luminescence evoked in 4 h by Tg (1 µM) and PMA (100 nM) in the indicated cell lines. Data are mean ± SEM of 8–12 biological replicates from three independent experiments. ( E ) Endogenous NFATC1 translocation evoked in 4 hr by Tg (1 µM) in the indicated cell lines, measured by immunofluorescence. Data are mean ± SEM of the nuclear to cytosol NFATC1 intensity ratio of 58–161 cells from four independent experiments. ( F ) Time-course of NFATC1 translocation evoked by plates coated with CD3 (OKT3 1 µg/ml). Data are mean ± SEM of 86–161 cells from five independent experiments. ( G ) IL-2 production evoked by untreated (NT) or surface coated CD3 (OKT3 1 µg/ml). Left, Representative density dot plots of Jurkat cells stained for IL-2. Graph bars represent the mean ± SEM of three independent experiments. One-way ANOVA Dunnett’s multiple comparisons test ( A ), Sidak multiple comparisons test ( D and E ), two-tailed unpaired Student’s t -test (C and G).
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Image Search Results


( A ) Averaged fura-2 responses, their peak amplitude (middle graph bar) and slope (right graph bar) evoked by Ca 2+ re-addition in Tg treated (1 µM, 8 min) Jurkat cells lines generated by CRISPR with control or ORAI1-targeted guiding sequences and stably re-expressing either WT or C143A ORAI1-YFP. Data are mean ± SEM of 210 (Control), 242 (KO), 189 (WT), and 203 (C143A) cells from three independent experiments ( B ) Western blot showing the amount of biotinylated GFP immunoreactivity in the PM in Jurkat CRISPR ORAI1 cells reconstituted with WT or C143A ORAI1-YFP. Representative of 2 independent experiments. ( C ) Individual (thin lines) and averaged (thick line) fura-2 recordings of Jurkat CRISPR ORAI1 cells reconstituted with WT or C143A ORAI1-YFP, exposed to CD3/CD28-coated beads in Ca 2+ containing solution (left). Averaged peak and integrated responses evoked by CD3/CD28 beads in individual cells during the recording period (right). Data are from 52 cells (WT) and 72 cells (C143A) from three independent experiments. ( D ) Relative changes in NFATC-Luciferase vs. housekeeping- Renilla luminescence evoked in 4 h by Tg (1 µM) and PMA (100 nM) in the indicated cell lines. Data are mean ± SEM of 8–12 biological replicates from three independent experiments. ( E ) Endogenous NFATC1 translocation evoked in 4 hr by Tg (1 µM) in the indicated cell lines, measured by immunofluorescence. Data are mean ± SEM of the nuclear to cytosol NFATC1 intensity ratio of 58–161 cells from four independent experiments. ( F ) Time-course of NFATC1 translocation evoked by plates coated with CD3 (OKT3 1 µg/ml). Data are mean ± SEM of 86–161 cells from five independent experiments. ( G ) IL-2 production evoked by untreated (NT) or surface coated CD3 (OKT3 1 µg/ml). Left, Representative density dot plots of Jurkat cells stained for IL-2. Graph bars represent the mean ± SEM of three independent experiments. One-way ANOVA Dunnett’s multiple comparisons test ( A ), Sidak multiple comparisons test ( D and E ), two-tailed unpaired Student’s t -test (C and G).

Journal: eLife

Article Title: S-acylation by ZDHHC20 targets ORAI1 channels to lipid rafts for efficient Ca 2+ signaling by Jurkat T cell receptors at the immune synapse

doi: 10.7554/eLife.72051

Figure Lengend Snippet: ( A ) Averaged fura-2 responses, their peak amplitude (middle graph bar) and slope (right graph bar) evoked by Ca 2+ re-addition in Tg treated (1 µM, 8 min) Jurkat cells lines generated by CRISPR with control or ORAI1-targeted guiding sequences and stably re-expressing either WT or C143A ORAI1-YFP. Data are mean ± SEM of 210 (Control), 242 (KO), 189 (WT), and 203 (C143A) cells from three independent experiments ( B ) Western blot showing the amount of biotinylated GFP immunoreactivity in the PM in Jurkat CRISPR ORAI1 cells reconstituted with WT or C143A ORAI1-YFP. Representative of 2 independent experiments. ( C ) Individual (thin lines) and averaged (thick line) fura-2 recordings of Jurkat CRISPR ORAI1 cells reconstituted with WT or C143A ORAI1-YFP, exposed to CD3/CD28-coated beads in Ca 2+ containing solution (left). Averaged peak and integrated responses evoked by CD3/CD28 beads in individual cells during the recording period (right). Data are from 52 cells (WT) and 72 cells (C143A) from three independent experiments. ( D ) Relative changes in NFATC-Luciferase vs. housekeeping- Renilla luminescence evoked in 4 h by Tg (1 µM) and PMA (100 nM) in the indicated cell lines. Data are mean ± SEM of 8–12 biological replicates from three independent experiments. ( E ) Endogenous NFATC1 translocation evoked in 4 hr by Tg (1 µM) in the indicated cell lines, measured by immunofluorescence. Data are mean ± SEM of the nuclear to cytosol NFATC1 intensity ratio of 58–161 cells from four independent experiments. ( F ) Time-course of NFATC1 translocation evoked by plates coated with CD3 (OKT3 1 µg/ml). Data are mean ± SEM of 86–161 cells from five independent experiments. ( G ) IL-2 production evoked by untreated (NT) or surface coated CD3 (OKT3 1 µg/ml). Left, Representative density dot plots of Jurkat cells stained for IL-2. Graph bars represent the mean ± SEM of three independent experiments. One-way ANOVA Dunnett’s multiple comparisons test ( A ), Sidak multiple comparisons test ( D and E ), two-tailed unpaired Student’s t -test (C and G).

Article Snippet: A total of 2 × 10 6 Jurkat cells were electroporated with Amaxa nucleofector (Kit V) with 1 µg of firefly luciferase encoding plasmid 9NFAT-luc together with 0.2 µg of control plasmid encoding the Renilla luciferase (phRL-TK-luc, Promega).

Techniques: Generated, CRISPR, Stable Transfection, Expressing, Western Blot, Luciferase, Translocation Assay, Immunofluorescence, Staining, Two Tailed Test

( A ) Conditions used to study synapse formation between Raji and Jurkat T cells. Raji are pulsed with SEE prior to co-culture with Jurkat cells, IS formation evaluated in living (25 min) and fixed cells (1 H) and Jurkat activation at 24 hr. ( B ) Relative changes in NFAT-Luciferase vs. housekeeping- Renilla luminescence evoked by coculture of the indicated cell lines for 24 hr with naïve or SEE (1 µg/ml) pulsed Raji. Data are mean ± SEM of 15–20 biological replicates in cells from four independent experiments. ( C ) Representative images of ORAI1-deficient Jurkat T cells reconstituted with WT or mutant ORAI1-YFP co-cultured with pulsed RAJI cells stained with CellMask deep red. (Scale bar = 10 µm). ( D ) ORAI distribution in IS vs. opposite pole in IS forming between SEE-pulsed Raji and ORAI1-deficient Jurkat T cells reconstituted with WT (29 IS) or mutant (26 IS) ORAI1. Chi-square p value: 0.0014, two-sided Fisher’s exact test. ( E ) Quantification of ORAI1 enrichment in time-lapse images from C. IS accumulation was measured using kymographs of 20 pixels wide lines spanning the IS-distal cap axis, drawn on stable (not moving) IS forming in WT (12) and C143A (8) cells from three independent experiments. Graph bar shows mean ± SEM area under the curve of the kinetic enrichment graphs. ( F ) Representative confocal images of IS forming between SEE-pulsed Raji labeled with CellMask (Magenta, APC white labelling) and ORAI1-deficient Jurkat T cells reconstituted with WT or mutant ORAI1-YFP (Green), labelled with Phalloidin (Blue) and anti-TCR-PE (Red). Scale bar = 10 µm. Arrows indicate IS formation and asterisks accumulation of ORAI1 in the opposite pole. ( G ) IS enrichment for ORAI1-YFP, Phalloidin and TCR fluorescence in Jurkat T cells reconstituted with WT or C143A ORAI1-YFP. Fluorescence levels at the IS and opposite pole were extracted from 10 pixel wide line profiles. Graph bars shows mean ± SEM in WT (n = 25 IS) and C143A (n = 20 IS). Two-tailed unpaired Student’s t -test ( B, E and G ); Chi square ( D ). ( H ) Proposed model: S-acylation by PAT20 targets ORAI1 to lipid rafts, enabling the coordinated recruitment of ORAI1 and TCR to the immune synapse for efficient signalling.

Journal: eLife

Article Title: S-acylation by ZDHHC20 targets ORAI1 channels to lipid rafts for efficient Ca 2+ signaling by Jurkat T cell receptors at the immune synapse

doi: 10.7554/eLife.72051

Figure Lengend Snippet: ( A ) Conditions used to study synapse formation between Raji and Jurkat T cells. Raji are pulsed with SEE prior to co-culture with Jurkat cells, IS formation evaluated in living (25 min) and fixed cells (1 H) and Jurkat activation at 24 hr. ( B ) Relative changes in NFAT-Luciferase vs. housekeeping- Renilla luminescence evoked by coculture of the indicated cell lines for 24 hr with naïve or SEE (1 µg/ml) pulsed Raji. Data are mean ± SEM of 15–20 biological replicates in cells from four independent experiments. ( C ) Representative images of ORAI1-deficient Jurkat T cells reconstituted with WT or mutant ORAI1-YFP co-cultured with pulsed RAJI cells stained with CellMask deep red. (Scale bar = 10 µm). ( D ) ORAI distribution in IS vs. opposite pole in IS forming between SEE-pulsed Raji and ORAI1-deficient Jurkat T cells reconstituted with WT (29 IS) or mutant (26 IS) ORAI1. Chi-square p value: 0.0014, two-sided Fisher’s exact test. ( E ) Quantification of ORAI1 enrichment in time-lapse images from C. IS accumulation was measured using kymographs of 20 pixels wide lines spanning the IS-distal cap axis, drawn on stable (not moving) IS forming in WT (12) and C143A (8) cells from three independent experiments. Graph bar shows mean ± SEM area under the curve of the kinetic enrichment graphs. ( F ) Representative confocal images of IS forming between SEE-pulsed Raji labeled with CellMask (Magenta, APC white labelling) and ORAI1-deficient Jurkat T cells reconstituted with WT or mutant ORAI1-YFP (Green), labelled with Phalloidin (Blue) and anti-TCR-PE (Red). Scale bar = 10 µm. Arrows indicate IS formation and asterisks accumulation of ORAI1 in the opposite pole. ( G ) IS enrichment for ORAI1-YFP, Phalloidin and TCR fluorescence in Jurkat T cells reconstituted with WT or C143A ORAI1-YFP. Fluorescence levels at the IS and opposite pole were extracted from 10 pixel wide line profiles. Graph bars shows mean ± SEM in WT (n = 25 IS) and C143A (n = 20 IS). Two-tailed unpaired Student’s t -test ( B, E and G ); Chi square ( D ). ( H ) Proposed model: S-acylation by PAT20 targets ORAI1 to lipid rafts, enabling the coordinated recruitment of ORAI1 and TCR to the immune synapse for efficient signalling.

Article Snippet: A total of 2 × 10 6 Jurkat cells were electroporated with Amaxa nucleofector (Kit V) with 1 µg of firefly luciferase encoding plasmid 9NFAT-luc together with 0.2 µg of control plasmid encoding the Renilla luciferase (phRL-TK-luc, Promega).

Techniques: Co-Culture Assay, Activation Assay, Luciferase, Mutagenesis, Cell Culture, Staining, Labeling, Fluorescence, Two Tailed Test